Verteporfin, photofrin II, and merocyanine 540 as PDT photosensitizers against melanoma cells

doi:10.1016/j.bbrc.2006.08.060

Biochemical and Biophysical Research Communications

Volume 349, Issue 2, 20 October 2006, Pages 549-555

https://doi.org/10.1016/j.bbrc.2006.08.060 Get rights and content

Abstract

The efficiency of photodynamic effect (PDE) for Photofrin II (PfII), Verteporfin, and Merocyanine 540 (MC540) was compared against neoplastic cells. Triplet state lifetimes and singlet molecular oxygen quantum yields were correlated with biological effect. PfII triplet lifetime was two times longer than that of Verteporfin, however, its singlet molecular oxygen quantum yield was two times lower in comparison with Verteporfin. High singlet molecular oxygen quantum yield of Verteporfin resulted in high biological efficacy. To achieve 50% mortality of cells four times lower light dose and five times lower concentration of Verteporfin were applied in comparison with PfII. The same level of cell damage was reached using 10 times higher light dose and two times higher concentration of MC540 in comparison with PfII. Our results confirm that singlet molecular oxygen based mechanism, prevalent for Verteporfin and PfII, was highly effective against melanoma cells. Verteporfin can be used at small doses with high cellular damage efficiency.

Section snippets

Materials and methods

Materials. Merocyanine 540 was purchased from Sigma and was used without any further purification. A stock solution was obtained by dissolving MC540 in DMSO and diluting the solution in phosphate-buffered saline without Ca²⁺ and Mg²⁺ (DMSO:PBS, 1:19). Photofrin II (Quadra Logic Technologies, Holland) containing ⩽11% of ester was diluted in 5% glucose. Visudyne^® (Verteporfin, BPD-MA) was the generous gift of Novartis Pharma and was used without any further purification. Aliquots of Verteporfin

Photostability of compounds

Exposure of PfII and Verteporfin to light dose employed in PDE studies (0.5 J/cm²) caused a very limited photodegradation. There was no significant change in absorbance at light dose tested (2% and 3% for PfII and Verteporfin, respectively). MC540 exhibited a slightly higher (up to 5%) decrease in absorbance.

Cytotoxicity assay

Toxicity of all photosensitizers against S91 melanoma cells was tested in dark (Fig. 1, insets). Concentration–response experiments were performed to establish the optimal dosage of

Discussion

The principal goal of the present study was to compare the efficiency of photodynamic effect using cyanine and porphyrin derivative photosensitizers against melanoma cells. Concentration of photosensitizer, light dose used, and mechanism of photosensitizer action are factors determining the level of biological damage.

Verteporfin turned out to be most potent photosensitizer against melanoma cells. The amount of the dye in cell culture can be 5- to 10-fold lower than in the case of PfII and

Acknowledgments

The authors gratefully acknowledge Novartis for delivering Visudyne^® and Professor Tadeusz Sarna for availability of singlet molecular oxygen emission intensity measurements. We thank Dr. Bozena Romanowska-Dixon for her help and support. We gratefully acknowledge Dr. Wojciech Macyk for power light measurements. This work was supported by the State Committee for Scientific Research (KBN, Poland) Grant No. 3 T09A 073 27. A preliminary report was presented at the Inorganic Reaction Mechanism

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^☆: Abbreviations: BPD-MA, benzoporphyrin derivative monoacid A; DMSO, dimethyl sulfoxide; FCS, fetal calf serum; ICG, Indocyanine green; LD_10,50,90, dose causing 10%, 50%, and 90% level of cell survival; M14, human melanoma cell line; MC540, Merocyanine 540; PBS, phosphate-buffered saline; PDE, photodynamic effect; PDT, photodynamic therapy; PfII, Photofrin II; PS, photosensitizer; RPMI 1640, cell culture medium; S91/I3, Cloudman mouse melanoma cell line; SKMEL 188, human melanoma cell line.

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Verteporfin, photofrin II, and merocyanine 540 as PDT photosensitizers against melanoma cells☆

Abstract

Section snippets

Materials and methods

Photostability of compounds

Cytotoxicity assay

Discussion

Acknowledgments

J. Photochem. Photobiol. B

Int. J. Radiat. Oncol. Biol. Phys.

J. Photochem. Photobiol. B

J. Photochem. Photobiol. B

J. Photochem. Photobiol. B

Biochim. Biophys. Acta

Chem. Phys.

Toxicol. Lett.

J. Photochem. Photobiol. B

Surv. Ophthalmol.

J. Photochem. Photobiol. A

J. Photochem. Photobiol. B

J. Photochem. Photobiol. B

Photodynamic therapeutics: basic principles and clinical applications

DDT

Rate of oxygen consumption of hamster melanoma cells as a factor influencing their radioresistance

Folia Histochem. Cytochem.

Effect of melanin on radiation response of CHO cells

Int. J. Radiat. Biol. Relat. Stud. Phys. Chem. Med.

Growth and pigmentation in genetically related Cloudman S91 melanoma cell lines treated with 3-isobutyl-1-methyl-xanthine and β-melanocyte-stimulating hormone

Exp. Dermatol.